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Detection of Citrus tatter leaf virus with reverse transcription—polymerase chain reaction (RT-PCR)

Detection of Citrus tatter leaf virus with reverse transcription—polymerase chain reaction (RT-PCR) Citrus tatter leaf virus (CTLV) has the potential to cause major losses to the Australian citrus industry if an infected clone is propagated, because the predominant rootstocks are intolerant of CTLV infection. We have developed a robust and specific semi-nested reverse transcription—polymerase chain reaction (RT-PCR) assay which detects CTLV in a range of citrus tissues. The sensitivity of the assay is at least 500 times greater than that of ELISA-based methods and allows detection directly from field trees. We have combined the assay with a simple and rapid tissue extraction protocol to make it amenable to large-scale screening. This method will improve the rapidity and reliability of CTLV screening of trees in the Australian Citrus Budwood Scheme. Nucleotide sequence analysis of the amplified CTLV fragments shows near identity (99.8%) amongst Australian isolates and between Australian isolates and a Japanese isolate of apple stem grooving virus (98.1%), and a high level of identity (92.0%) to a Japanese isolate of CTLV. http://www.deepdyve.com/assets/images/DeepDyve-Logo-lg.png Australasian Plant Pathology Springer Journals

Detection of Citrus tatter leaf virus with reverse transcription—polymerase chain reaction (RT-PCR)

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References (20)

Publisher
Springer Journals
Copyright
Copyright © 2000 by Australasian Plant Pathology Society
Subject
Life Sciences; Plant Pathology; Plant Sciences; Agriculture; Entomology; Ecology
ISSN
0815-3191
eISSN
1448-6032
DOI
10.1071/AP00046
Publisher site
See Article on Publisher Site

Abstract

Citrus tatter leaf virus (CTLV) has the potential to cause major losses to the Australian citrus industry if an infected clone is propagated, because the predominant rootstocks are intolerant of CTLV infection. We have developed a robust and specific semi-nested reverse transcription—polymerase chain reaction (RT-PCR) assay which detects CTLV in a range of citrus tissues. The sensitivity of the assay is at least 500 times greater than that of ELISA-based methods and allows detection directly from field trees. We have combined the assay with a simple and rapid tissue extraction protocol to make it amenable to large-scale screening. This method will improve the rapidity and reliability of CTLV screening of trees in the Australian Citrus Budwood Scheme. Nucleotide sequence analysis of the amplified CTLV fragments shows near identity (99.8%) amongst Australian isolates and between Australian isolates and a Japanese isolate of apple stem grooving virus (98.1%), and a high level of identity (92.0%) to a Japanese isolate of CTLV.

Journal

Australasian Plant PathologySpringer Journals

Published: Jan 28, 2011

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